醫(yī)學(xué)-以bcl2蛋白家族為靶點(diǎn)的抗腫瘤藥物設(shè)計(jì)

以BCL-2蛋白家族為靶點(diǎn)的抗腫瘤藥物設(shè)計(jì)Bcl-2FamilyProteinsasTherapeuticTargetsforanticancerdrugdesignDate:2021-05-231OutlineBackgroundHallmarksofCancerTheBcl-2ProteinFamilyModelofBH3mimetic-inducedapoptosisStructurebasisfordesigningligandsSmallmoleculeinhibitorsPeptidomimeticapproach—mimicthea-helixoftheBH3peptidesABT-737,ABT-263andtheiranalogspan-activeinhibitorofBcl-2SelectiveBcl-2inhibitorsselectiveBCL-xLinhibitorSelectiveMcl-1inhibitors2,Cell,144(5):646-674HallmarksofCancer3ExtrinsicandintrinsicpathwaysofapoptosisExpertOpin.Ther.Patents(2021),22(1):37-554TheBcl-2ProteinFamilyTheBcl-2FamilyProteinswithanti-orpro-apoptoticfunction

Characteristics:(Often)aC-terminaltransmembranedomain.Bcl-2homology(BH)domains.Bcl-2proteinsformhomo-andheterodimersviatheirBH-domainsBH1,BH2: PredictedtoformionchannelsBH3: The〞suicidedomain〞–regulatescelldeathBH4: Thoughttoconferanti-apoptoticactivity.(Chan&Yu.(2004).Clin.Exp.Pharmacol.Physiol.31:119).Chipuk,J.E，MolecularCell，2010，37(3):299-3105ElevatedofBcl-2orBcl-xLexpressionSeminarsinOncology,2004,30(Supplement16):133-142.6ModelofBH3mimetic-inducedapoptosis一、直接模型：

抑制劑代替促凋亡蛋白BAK/BAX，與抗凋亡蛋白Bcl-2/Bcl-xl結(jié)合，從而抑制后者，促進(jìn)細(xì)胞凋亡二、間接模型

抑制劑替代特定的激活因子BH3-only蛋白，與抗調(diào)亡蛋白如BCL-2/BCL-xL結(jié)合，游離出來(lái)的BH3-only蛋白，激活BAK/BAX。NatureReviewsDrugDiscovery2021，7(12):989-1000ClinicalCancerResearch2021，15(4):1126-1132.Oncogene，2021，27(S1):S149-S157.7Structurebasisfordesigningligands8ThemolecularbasisforheterodimerformationHeterodimerizationbetweenmembersoftheBcl-2familyofproteinsisakeyeventintheregulationofapoptosispro-apoptoticprotein(Bak)adoptsanamphipathica-helix(BH3domain)thatinteractswithanti-apoptoticproteins(Bcl-xl)throughhydrophobicandelectrostaticinteractionsinthehydrophobiccleft.9抗凋亡蛋白(BCL-2)是通過(guò)其外表疏水凹槽與促凋亡類(BIM)的BH3區(qū)域結(jié)合發(fā)生相互作用，調(diào)節(jié)細(xì)胞正常的生理凋亡。BCL-2familyantagonistsforcancertherapy.NatureReviewsDrugDiscovery2021，7(12):989-1000OverlayofBCL-XL–ABT-737complexwithBCL-xlL–BIMBH3complexBcl-2andBIMBH310Bcl-xLandBak-BH3SurfacerepresentationofthebindingpocketofBcl-xLboundtotheBakpeptide11."StructureofBcl-xL-BakPeptideComplex:RecognitionBetweenRegulatorsofApoptosis."Science1997，275(5302):983-986.Bcl-xL----BimBH3Mcl-1—BimBH3Bcl-xL/BadProteinScience,2000,9:2528–2534125個(gè)疏水中心

(P1,P2,P3,P4和P5)4個(gè)氫鍵供體(D1,D2,D3和D4)2個(gè)氫鍵受體(A1andA2)Bcl-2活性部位分布圖在受體D1、D2、D3、A1位點(diǎn)出現(xiàn)比較頻繁，與受體形成靜電作用能提高親和力與特異性D4，A2位點(diǎn)出現(xiàn)頻率較小，能量奉獻(xiàn)不大BMC，2007，15(19):6407-6417SmallmoleculeinhibitorsPeptidomimeticapproach—mimicthea-helixoftheBH3peptidesABT-737,ABT-263andtheiranalogspan-activeinhibitorofBcl-2SelectiveBcl-2inhibitorsselectiveBCL-xLinhibitorSelectiveMcl-1inhibitors14BakPeptidomimeticapproach—mimicthea-helixoftheBH3peptides

JACS,2005,127(29):10191-10196JACS,2005,127(15):5,463-546815JBiolChem.2021,18;286(11):9382–9392OrganicLetters,2007,9(19):3733-3736.BimBH3a-helixJ.Med.Chem.2021,55,10735?1074116ABT-737,ABT-263andtheiranalogs17SARbyNMR與ABT-737系列的發(fā)現(xiàn)3.Screenforsecondligand2D1H-15NHSQC譜中蛋白的H1或N15化學(xué)位移變化。18Science.1996Nov29;274(5292):1531-4.4.基于上述三維結(jié)構(gòu)設(shè)計(jì)恰當(dāng)?shù)倪B接橋?qū)蓚€(gè)片段連接起來(lái)，使得到的分子和靶蛋白結(jié)合時(shí)保持各自獨(dú)立時(shí)的結(jié)合位置及其空間取向，最終篩選得到一個(gè)高親和性的配體.特點(diǎn)：首先篩選結(jié)合于生物靶分子亞活性位點(diǎn)的低親和性配體，然后通過(guò)優(yōu)化和組裝便得到所期望的高親和性配體。優(yōu)點(diǎn)：NMR實(shí)驗(yàn)方法來(lái)區(qū)分小分子和靶蛋白的結(jié)合，能快速地從小分子化合物庫(kù)中發(fā)現(xiàn)結(jié)合于靶蛋白亞活性位點(diǎn)的低親和性配體。如Fesik等篩選溶基質(zhì)素(stromelysin)的抑制劑只用了半年時(shí)間。限制：知道靶蛋白確切的NMR三維結(jié)構(gòu)、提供足夠量的15N標(biāo)記的靶蛋白(>200mg)〔<25kDa〕、解析蛋白質(zhì)三維結(jié)構(gòu)的能力。Linkligands連接基團(tuán)成功的關(guān)鍵是連接橋的長(zhǎng)度及其性質(zhì)〔如剛性〕Science274,1531-1534(1996)19ABT-737系列的發(fā)現(xiàn)過(guò)程1.為BCL-xL篩選第一個(gè)配體。從平均分子量為200Da的10000個(gè)化合物片段庫(kù)中篩選出來(lái)1黑：HSQCof15N-Bcl-xL紅：HSQCof15N-Bcl-xLwithbiarylacid〔1〕1的羧基能與R139結(jié)合，相當(dāng)與Bcl-Xl—Bak復(fù)合物中的D8320Site1:Tyr101,Leu108,Val126andPhe146212.篩選第二個(gè)配體從平均分子量為125Da的3500個(gè)化合物中篩選出11黑:HSQCof15N-Bcl-xL.紅:HSQCof15N-Bcl-xLwithbiarylacid(1).綠:HSQCof15N-Bcl-xLwithnapthol(11).22

連接方式25的活性比1提高了大概200倍，但溶解度不好1123JMedChem49,656-663(2006)Site224A-38535831不幸的是，31在HSA存在情況下與Bcl-Xl結(jié)合大大下降。它能與HSA的domainIII緊密結(jié)合。J.Med.Chem.2006,49,1165-118168倍的失活Bcl-xLKi<1nMBcl-xLKi10%HS:35nM25增加site1的極性ABT-263-----口服小分子抑制劑ABT-737ABT-263MW:974Pka10J.Med.Chem.2021,51,6902–6915NatureReviewsDrugDiscovery2021，7(12):989-100026MW：813.43ABT-737/ABT-263類似物〔以結(jié)構(gòu)為根底的藥物設(shè)計(jì)〕喹唑啉類JACS.2006,128,16206-16212J.Med.Chem.2021,54,1914–1926J.Med.Chem.2021,55,4664?4682J.Med.Chem.2021,55,6149?6161J.Med.Chem.2021,55,8502?8514J.Med.Chem.2021,56,3048?30674,5-二苯基吡唑類Bcl-2/Bcl-xL:Ki<1nMAOptimizationBM-9572728總結(jié)1、ABT-737/ABT-263打破了lipinsk’sruleoffive。一方面是由于雅培公司研究人員用SARbyNMR，平行合成，基于結(jié)構(gòu)藥物設(shè)計(jì)多種方式設(shè)計(jì)小分子抑制劑；另一方面，對(duì)小分子與受體結(jié)合模式的進(jìn)行雙向研究，相互理解。2、對(duì)研究者研究Bcl-2家族蛋白，及設(shè)計(jì)小分子抑制劑有很好的指導(dǎo)作用：BCl-2/BCl-Xl三維結(jié)構(gòu)的理解、SARbyNMR、Structure-BasedDesign，F(xiàn)BDD等。29pan-activeinhibitorofBcl-230ObatoclaxPhaseIIAT-101PhaseII/III早期研發(fā)的抗凋亡蛋白小分子抑制劑根本上都是pan-Bcl-2antagonist包括三聯(lián)苯類，HA-14-1及其類似物，棉酚及其衍生物，BH3I及其衍生物等

BH3I-1S1DCBL55JournalofMedicinalChemistry,2021,53(9):3465-3479NatureReviewsDrugDiscovery,2021,7(12):989-100031ABT-199SelectivelyKillsBCL-2-DependentTumorCellsWhileSparingPlatelets(A)Navitoclax(ABT-263)bindswithhighaffinitytobothBCL-2andBCL-XL.Becausemanytumors,particularlylymphoidmalignancies,areaddictedtoBCL-2forsurvival,...CancerCell,2021,23(2),139-141ABT-199：SelectiveBcl-2inhibitor32Naturemedicine,2021,19(2):202-8BCL-2Ki<0.010nMBCL-XLKi=48nMBCL-WKi=245nMPhaseI/IIPhaseI33化合物1與Bcl-2化合物2與Bcl-234其它Y